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Bioremediation As a Tool for Resiliency in Oil Spills

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Bioremediation As a Tool for Resiliency in Oil Spills Bioremediation as a Tool for Resiliency in Oil Spills By: Caitlin G. Bergman, Elena R. Eberhart, Mackenzie A. Garner, Atticus H. Hamilton, and Claire M. Bredar Coach: Chris Backstrum South Anchorage High School 13400 Elmore Road Anchorage, Alaska 99504 Contact person: Caitlin Bergman at [email protected] “This paper was written as part of the Alaska Ocean Sciences Bowl high school competition. The conclusions in this report are solely those of the student authors.” Bioremediation as a Tool for Resiliency in Oil Spills Abstract With an increase in boating, offshore drilling, and transportation of oil, coastal communities are in need of a plan to clean up waterways in the event of an offshore oil spill. Manual removal of oil is a critical first step and when paired with bioremediation as a secondary method the highest possible success of oil breakdown at a spill site can be achieved. Unlike dispersants, bioremediation is a clean way to speed up the degeneration of oil in waterways. According to research done by Gordon in 1994, given that proper nutrients are present, an oil spill that was estimated to be cleaned by natural conditions in 5-10 years could be cleaned in 2-5 years with the use of bioremediation. Alcanivorax borkumensis, an oil-degrading microbe, was added to water with heavy fuel oil. After two months the water was clean enough to return back into the seas (Golyshin, n.d.). This bacteria is found in marine ecosystems and can absorb and digest linear and branched alkanes that are found in crude oil and its products (Rojo, 2009). Community resilience to oil spills would involve establishing a preparedness plan with a system in place to quickly implement bioremediation methods. Establishing oil spill preparedness practices in coastal communities will be critical to reduce impact from oil. This system would incorporate culturing tanks, quantities of hydrocarbonoclastic bacteria in pellet form, and the means to transport and apply in the field. Introduction When oil spills occur, they can cause major damage to the environment. There have been many oil spills in Alaska (Table 1), with devastating environmental effects. Many marine mammals have been killed, fisheries have been closed, and native populations have lost their food source as a result of oil spills (Amadeo, 2015). It is very important for communities to have a plan for what to do in the event of an oil spill. Bioremediation uses oil-degrading bacteria to clean up the spill. Bioremediation has already been used in many oil spills, including the spill in Prince William Sound. Adding bacteria and nutrients to oil spills will significantly increase the degradation of the oil, with less environmental impacts than other cleanup methods. This will help communities to be more resilient, and recover more quickly after the spill. Bioremediation Bioremediation is the use of naturally occurring or introduced microorganisms or other forms of life to consume or break down environmental pollution in order to clean up the polluted area (Collins English Dictionary, 2015). Bioremediation is an alternative cleanup action that is safer for the environment than other chemical or physical solutions. The microorganisms involved with bioremediation are either already living in the affected environment or they are brought into the environment (Cornell, 2009). If the chosen bioremediation process involves the naturally occurring microorganisms in the ocean environment, then the group organizing the bioremediation cleanup would add 1 nutrients to the water to help boost population growth of the native species of bacteria. This was used in Alaska, during the Exxon Valdez oil spill in Prince William Sound. Fertilizer was added to beaches in order to supply nutrients to naturally occurring microbes in the water. The oil in fertilized areas biodegraded about two times faster than untreated controls (Pritchard et al., 1992). The other main method of bioremediation is to take cultured microorganisms and introduce them into an oil spill and then continue to feed them, making the population grow. Composition of Crude Oil Crude oil is made up of four main elements. It usually contains 84% to 87% carbon, 11% to 14% hydrogen, 0.1% to 8% sulfur, 0.1% to 1.8% nitrogen, and 1% to 1.5% oxygen (Lyons, 2005). Although there are sodium, nitrogen, and oxygen compounds in oil, the most common molecules are hydrocarbons. There are three main groups of hydrocarbon molecules in crude oil; aromatics, naphthenes, and alkanes (Australian Institute of Petroleum, 2013). The most difficult hydrocarbons for bacteria to biodegrade are aromatic compounds. Aromatic compounds are double-bonded carbon rings. Some of them also have attached chains of hydrocarbons. Very small (one or two ring) aromatics evaporate off of a spill or can be biodegraded. Larger aromatics, however, resist biodegradation, and can persist in the area of a spill for a long time (American Academy of Microbiology, 2011). The only way that they can be broken down is by photo oxidation, or degradation by U.V. light. Aromatics are also the most toxic compounds in crude oil (NOAA, 2015). 2 Naphthenes are single-bonded, saturated hydrocarbon rings. Naphthenes can be biodegraded more easily than aromatics, but not as quickly as alkanes because they contain more bonds (El-Nemr, 2006). Alkanes are straight or branched saturated hydrocarbons. They only contain single bonds, which is ideal for microbial degradation because it does not take much energy to break apart the molecules, compared to double-bonded molecules. They can be solids, liquids, or gases, depending on the number of carbon atoms they contain. Alkanes with one to four carbon atoms are gases, also called volatile compounds. During an oil spill, these compounds evaporate off of the slick and into the air. Alkanes with five to sixteen carbon atoms are liquids. These form most of the oil slick. They can be degraded relatively quickly by bacteria. The smaller the chain, the easier it is for bacteria to break it down. Alkanes with more that sixteen carbon atoms are solids, and are difficult for bacteria to break down. The characteristics of crude oil differ from place to place. Alaskan North Slope crude oil has a relatively high viscosity, and forms an emulsion with water very quickly (NOAA, 2015). This makes it extremely important to respond quickly to spills made up of this type of oil. With the exception of aromatic compounds, all hydrocarbons in crude oil can be degraded by bacteria. This makes bioremediation a very important method for cleaning up oil spills. In time, naturally occurring bacteria will completely break down a spill. However, they cannot do this quickly enough to prevent damage to the ecosystem, unless bioremediation methods are used to speed up their growth. 3 Alcanivorax borkumensis Alcanivorax borkumensis is a marine bacteria that can absorb and digest linear and branched alkanes that are found in crude oil and its products (Rojo, 2009). A. borkumensis is a gram-negative bacteria, meaning that the bacteria has an outer membrane of lipopolysaccharides, unlike gram positive bacteria who do not possess this layer. It is also a rod-shaped bacterium that is aerobic (oxygen reliant) (Biello, 2006). A. borkumensis is included in the genus Bacillus, which is a genus for rod-shaped bacterium and is in the class Gammaproteobacteria, meaning that it is a scientifically important bacteria (Kostka et al., 2011). Since A. borkumensis occurs naturally in unpolluted waters all over the world (including freshwater), it has to have a source of energy. A particular study has found that strains of two of the most abundant cyanobacteria in the ocean (Prochlorococcus and Synechococcus) produce and accumulate hydrocarbons, particularly alkanes C15 and C17 (PNAS, n.d.). These alkanes are the energy source for A. borkumensis in unpolluted water. A. borkumensis naturally flourishes after an oil spill because there is a more abundant source of energy that can sustain a larger population (Kimes, 2014). A. borkumensis also participates in wastewater treatment by being foamed by Nocardia spp. (Shamoon, n.d). A borkumensis breaks apart the bonds in hydrocarbons in oil that have been exposed to the sea, using enzymes and oxygen found in the seawater ( Biello, 2010). A. borkumensis creates enzymes AlkB1 and AlkB2 (Beilen, 2004). AlkB1 is involved with the direct reversal of alkylation damage, specifically in single-stranded DNA ( Dinglay, 2000). AlkB1 hydroxylases alkanes with 5 to 12 carbons, and AlkB2 hydroxylases alkanes with 8 to 16 carbons (Rojo, 2009). The chain lengths with the most 4 A. borkumensis growth are 14 to 19 carbon chains (Naether et al., 2013). A. borkumensis is able to outcompete other hydrocarbonoclastic species of bacteria because it can break down such a wide range of alkane chains (Hara, Akihiro, Kazuaki Syutsubo, & Shigeaki Harayama, 2003). A. borkumensis cannot consume sugars or amino acids as a source of energy, unlike most other bacteria (Yakimov, Michail M., et al, 1998). A. borkumensis Genetics The Alcanivorax Borkumensis bacterium contains a single ringed chromosome free floating in the cytoplasm. This single stranded circular chromosome contains islands. It is these islands that code for the bacteria’s oil degrading enzymes. The islands are made up of genes that code for different enzymes. These genes are mobile, meaning that they can change their position in the chromosome, also allowing the islands to transfer to other bacteria of different species through horizontal gene transfer. In this way, non-oil degrading microbes can develop the enzyme creation by horizontal gene transfer with A. borkumensis. Then, they are able to degrade oil. Alcanivorax borkumensis has in its SK2 genome a number of regions, and one such island contains a cluster of 40 genes which code for cell surface biosynthesis. The second island holds a complete cluster of genes for alkane degradation.
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